LOCUS       sequence-120427-        5074 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PSmOrange targeted to mitochondria using the synthetic LOC8R 
            presequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5074)
  TITLE     Mitochondrial LOC8R Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5074)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: LOC8R-PSmOrange
FEATURES             Location/Qualifiers
     source          1..5074
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             634..708
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             730..804
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             826..900
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             922..996
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     regulatory      1024..1033
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1030..1740
                     /codon_start=1
                     /product="orange to far-red photoswitchable variant of the 
                     monomeric fluorescent protein mOrange (Subach et al., 
                     2011)"
                     /label=PSmOrange
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPLFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNY
                     EDGGVVTVTQDSSLQDGEFIYKVKMRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIRMRLKLKDGGHYTSEVKTTYKAKKSVQLPGAYIVGIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     1637..1660
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    1862..1983
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1899..1918)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1953..1972
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1990..2445)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2127..2148)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2339..2358
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2472..2576
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2578..2935
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2578..2769
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2601..2621)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2786..2921
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2970..3764
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2970..3764
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3024..3043)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3634..3653
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3952..3971)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3996..4043
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4372..4960
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4861..4880
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttgcc accatgagcg tgctgacccc cctgctgctg
      661 agaggcctga caggctctgc tagaaggctg cctgtgcctc gagccaagat ccacagcctg
      721 ggggatccaa tgtccgtgct gacacctctg ctgctgcggg gactgaccgg aagtgctcgg
      781 agactgccag tgcccagggc caagattcac tctctggggg atcccatgag tgtgctgact
      841 ccactgctgc tgcgcggcct gactggatca gcacgccgcc tgccagtgcc tcgagctaag
      901 attcattccc tgggcgaccc tatgtctgtg ctgacccctc tgctgctgag ggggctgact
      961 ggatctgcaa ggcggctgcc tgtgccaagg gccaaaatcc atagtctggg cgacccaccg
     1021 gtcgccacca tggtgagcaa gggcgaggag aataatatgg ccatcatcaa ggagttcatg
     1081 cgcttcaagg tgcgcatgga gggcaccgtg aacggccacg agttcgagat cgagggcgag
     1141 ggcgagggcc gcccctacga gggctttcag accgctaagc tgaaggtgac caagggcggc
     1201 cccctgccct tcgcctggga catcctgtcc cctctcttca cctacggctc caaggcctac
     1261 gtgaagcacc ccgccgacat ccccgactac ttcaagctgt ccttccccga gggcttcaag
     1321 tgggagcgcg tgatgaacta cgaggacggc ggcgtggtga ccgtgaccca ggactcctca
     1381 ctgcaggacg gcgagttcat ctacaaggtg aagatgcgcg gcaccaactt cccctccgac
     1441 ggccccgtga tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc
     1501 gaggacggcg ccctgaaggg cgagatcagg atgaggctga agctgaagga cggcggccac
     1561 tacacctccg aggtcaagac cacctacaag gccaagaagt ccgtgcagct gcccggcgcc
     1621 tacatcgtcg gcatcaagct ggacatcacc tcccacaacg aggactacac catcgtggaa
     1681 cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtaa
     1741 gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt
     1801 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     1861 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     1921 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     1981 ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     2041 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     2101 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     2161 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     2221 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     2281 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     2341 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     2401 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     2461 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     2521 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg
     2581 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     2641 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     2701 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     2761 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     2821 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     2881 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg
     2941 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     3001 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     3061 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     3121 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg
     3181 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     3241 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     3301 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     3361 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     3421 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     3481 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     3541 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     3601 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     3661 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     3721 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     3781 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     3841 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     3901 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg
     3961 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     4021 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     4081 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     4141 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     4201 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
     4261 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     4321 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4381 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4441 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4501 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4561 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4621 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4681 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4741 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4801 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4861 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4921 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4981 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     5041 tgattctgtg gataaccgta ttaccgccat gcat
//
